Abstract:In this study, 2-(4-fluorophenyl)-4,7-di(thiophene-2-yl)-1H-benzo[d]imidazole, a novel monomer, was synthesized and its polymer was used for the immobilization matrix for cholesterol oxidase (ChOx). A novel cholesterol biosensor was generated. The designed conducting polymer based biosensor showed good KMapp, Imax values and very low limit of detection value with the durable shelf life. Moreover, this electrochemical biosensor system will be functionalized with gold nanoparticles (AuNPs) to achieve more sensitive and rapid biosensors with high electron transfer ability of AuNPs.